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1

Separation of ampicillin on polar-endcapped phase : Development of the HPLC method to achieve its correct dosage in cardiac surgery

Kubickova Vendula, Racova Zuzana, Strojil Jan, Santavy Petr, Urbanek Karel

Acta Chromatographica

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2023

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Vol. 35, no. 1

1--9

EN

The accurate, simple, and selective reversed phase high performance liquid chromatography (RP-HPLC) has been established and validated for the determination of an antibiotic ampicillin (AMP) in human blood plasma. The SPE extraction was used for the sample preparation. Chromatographic separation was accomplished by a mobile phase containing 15 mM monopotassium phosphate solution of pH 3.3 and methanol (75:25, v/v) in an isocratic mode at a flow rate of 1.4 mL min⁻¹ at 30 °C. The separation was evaluated on a column with a new polar-endcapped C18 stationary phase Arion® Polar C18 or well-known phase Luna® Omega Polar C18. Excellent linearity (R2 0.9998) was shown over range 10–300 mg L⁻¹ with mean percentage recovery 90%. Peak shapes were symmetrical in both columns, Arion® Polar C18 and Luna® Omega Polar C18, with asymmetry factor of 1.0 and 1.4, tailing factor of 1.0 and 1.2, and retention factor of 4.6 and 5.6, respectively. The Arion® Polar C18 was almost 1.4-fold more effective than Luna® Omega Polar C18 phase. The LOQ for ampicillin was achieved 10 mg L⁻¹ for Luna® Omega Polar C18 and 5 mg L⁻¹ for Arion® Polar C18 using 20 µL of a solution containing 0.24 mg mL⁻¹ of cephalexin as an internal standard. A number of articles dealing with the determination of ampicillin is limited, therefore, this study showed the HPLC method suitable for the determination of AMP in human blood plasma from patient who underwent elective cardiac surgical revascularization. In addition, the determination of AMP was also performed for the first time using an Arion® Polar C18 column, which effectively separated AMP from other compounds present in human blood plasma. This new polar-endcapped phase can help in separation of polar antibiotics or other polar compounds, which are unsuccessfully separated on conventional C18 column, and thus can help during method development.

2

A validated new RP-HPLC method for simultaneous determination of amoxicillin, ampicillin and cloxacillin in pharmaceutical formulations

Gebretsadik Hailekiros, Kahsay Getu, Eticha Tadele, Gebretsadikan Tesfamichael

Acta Chromatographica

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2023

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Vol. 35, no. 2

193--203

EN

As per the World Health Organization, 10% of medicines in low- and middle-income nations are of poor quality and pose a huge public health risk. The development and implementation of cost-effective, efficient and quick analytical methods to control the quality of these medicines is one of the immediate strategies to avoid such a situation. Hence, the main goal of this study was to develop and validate a simple, specific and precise new RP–HPLC method for simultaneous analysis of amoxicillin, ampicillin and cloxacillin in pharmaceutical formulations. The chromatographic analysis was achieved using Shodex C18 (250 × 4.6 mm, 5 μm) column with UV detection at 225 nm. The mobile phase was a gradient mixture of 30 mM phosphate buffer, pH 4.0 (mobile phase A) and acetonitrile (mobile phase B). Efficient separation of the three drugs was obtained using the final optimized chromatographic conditions. The developed method was validated for its specificity, linearity, precision, accuracy and robustness as per the ICH guidelines. The validation results showed that the method was specific, linear, precise, accurate and robust for the simultaneous determination of the three drugs. The developed method was applied to determine the content of the three drugs in pharmaceutical formulations. The assay results of the preparations showed that their drug content was within the pharmacopeial limit stipulated for each drug product. It can be concluded that the proposed method is suitable for simultaneous determination of amoxicillin, ampicillin and cloxacillin in pharmaceutical formulations in industries and regulatory laboratories.

3

Development of Daphnia magna under exposure to ampicillin

Vo T.-M.-C., Pham N.-H., Nguyen T.-D., Bui M.-H., Dao T.-S.

Architecture Civil Engineering Environment

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2018

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Vol. 11, no. 3

147-152

EN

In this study, the chronic effects of Ampicillin on survival, reproduction and growth of Daphnia magna was monitored during 21 days exposure test. The results showed that Ampicillin strongly affected on life history traits of Daphnia magna, i.e. 47% reduction of survival proportion, lower 68% reproduction than the control, and reduction in body length and in the intrinsic population rate, especially at its highest concentration. The antibiotics especially Ampicillin should be included int the Vietnam guidelines for environmental and ecological protection.

4

Determination of ampicillin and amoxicillin by high-performance thin-layer chromatography

Ghoulipour V., Shokri M, Waqif-Husain S.

Acta Chromatographica

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2011

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Vol. 23, no. 3

483--498

EN

A simple and fast high-performance thin-layer chromatographic method has been developed for the simultaneous determination of ampicillin and amoxicillin. Titanium(IV) silicate (a synthetic inorganic ion-exchanger)-coated thin-layer chromatography (TLC) plates were used to separate them, employing a mixture of K2HPO4 (0.1 M) + KH2PO4 (0.1 M), 1:1 (υ/υ), as mobile phase. The development time was 18 min. The plates were sprayed with fresh 1% solution of ninhydrin in ethanol. The developed method enables highly contrasted chromatograms with red purple spots in white background. Densitometric measurements were made at wavelength 546 nm using Camag TLC Scanner-3. The ampicillin and amoxicillin recovery of the total procedure were equal to 99.99 and 100.43, respectively. The procedure is quantitatively characterized. Linearities were r2 > 0.9958 and 0.9954 for ampicillin and amoxicillin, respectively, and the relative standard deviations were <0.89 and 0.61, respectively. The limits of detection were 2.9 and 1.5 ng per spot and the limits of quantification were 14.5 and 7.5 ng per spot, respectively. The method is rapid, selective, precise, and accurate and thus can be used for the routine analysis of pharmaceutical preparations in quality control laboratories of the pharmaceutical industry. The method is successfully applied for the determination of ampicillin and amoxicillin in human blood plasma and urine.

5

Fotokatalityczna degradacja ampicyliny w roztworach wodnych

Makowski A., Sobczak A., Wcisło D., Adamek E., Baran W., Nocoń W.

Proceedings of ECOpole

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2009

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Vol. 3, No. 1

81--86

PL

Metody zaawansowanego utleniania, polegające na generowaniu reaktywnych rodników hydroksylowych, są stosowane do usuwania antybiotyków z roztworów wodnych. Mogą być też stosowane do degradacji antybiotyków do związków przejściowych, które następnie ulegają łatwiej biodegradacji. Roztwory antybiotyków naświetlano promieniowaniem UV λ = 366 nm w obecności fotokatalizatora P 25 Degussa. Zawartość ampicyliny i jej produktów przejściowych w trakcie fotodegradacji badano metodą HPLC. Stałe szybkości rozkładu ampicyliny wyznaczano z zależności logC/Co od czasu trwania procesu. Dodatek jonów żelaza do roztworu ampicyliny powoduje nieznaczny spadek szybkości jej degradacji. Wpływ jonów miedzi zależy od stosunku molowego metalu do antybiotyku. Wpływ jonów wapnia na szybkość rozpadu ampicyliny jest niewielki, natomiast w przypadku jonów magnezu i jonów kobaltu powoduje spadek szybkości degradacji ampicyliny. Na powierzchni katalizatora P 25 adsorbuje się ampicylina; proces ulega zmianie w obecności jonów metali i jonów fosforanowych.

EN

The advanced oxidation methods are applied to remove antibiotics from water solutions. They involve generating hydroxylic radicals (.OH) with high oxidation potential, which are able to mineralize organic compounds. They can be use to degrade antibiotics to transient metabolites, which are then easily biodegradable. They were irradiated with UV λ = 366 nm in the presence of P 25 Degussa photocatalyst. The level of ampicillin and its transient products during photocatalytic degradation was tested using the HPLC method. To estimate the photocatalytic degradation rate constants of ampicillin, the relationship log C/Co from time of duration of photocatalytic process were defined. The addition of iron ions to the ampicillin solution slightly slows down its photocatalytic degradation. The influence of copper ions on the speed of ampicillin degradation depends on the molar ratio of metal to antibiotic. The influence of calcium ions on the speed of ampicillin breakdown is not too significant, while in the case of magnesium ions, similar to cobalt ions, the ampicillin breakdown slows down. Ampicillin is adsorbed on the surface of P 25 catalyst; the process changes in the presence of metal ions and phosphate ions. The optimum of environment during the photodegradation process is pH reaction 7.2.

6

Spectrophotometric determination of ampicillin and amoxycillin with Ce(IV) and Arsenazo III

Buhl F., Szpikowska-Sroka B.

Chemia Analityczna

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2001

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Vol. 46, No. 1

69-73

EN

A simple and sensitive spectrophotometric method for the determination of ampicillin and amoxycillin in a system: Ce(IV), reductant, arsenazo III was carried out. Colori-metric method for the determination of the analyte is based on reduction of cerium(IV) ions with ampicillin and amoxycillin. The cerium(III) ions formed were complexed with arsenazo III and spectrophotometrically determined. Beer's law is obeyed in the concentration range from 2.5 to 25ug and from 2 to 10 ng in 25ml for ampicillin and amoxycillin, respectively. The absorbance was measured at lambda(max) = 655 nm at pH 3.2-3.6 in 1cm cuvettes. The molar absorptivity of the colour system was found to be 2.7 x 10(5) 1 mo(-1)cm(-1) for ampicillin and 5.2 x 10(5)mol (-1)cm(-1) for amoxycillin. The method has been successfully applied for the determination of ampicillin and amoxycillin in pharma-ceuticals.

PL

Opracowano prostą i czułą metodę oznaczania ampicyliny i amoksycyliny w układzie Ce(IV), reduktor, arsenazo III. Kolorymetryczna metoda oznaczania analitów opiera się na redukcji Ce(IV) za pomocą ampicyliny i amoksycyliny, a następnie kompleksowania powstałych w stechiometrycznej ilości jonów Ce(III) przez arsenazo III. Pomiary przeprowadzono przy pH 3.2-3.6, Lambda max = 655 nm, w kuwetach o grubości l cm. Roztwory spełniają prawo Beera w zakresie od 2.5 do 25 ug w25 ml dla ampicyliny i od 2 do 10 ng w 25 ml dla amoksycyliny. Molowy współczynnik absorpcji liczony w stosunku do oznaczanych analitów wynosi 2.7 x 10(5)lmol(-1)cm(-1) dla ampicyliny i 5.2 * 10(5)1mol(-1)cm(-1) dla amoksycyliny. Metoda ta została zastosowana do oznaczania analitów w preparatach farmaceutyczny ch.

7

Kinetics and Mechanism of Oxidative transformations of (6R)-6-(A-Phenyl-D-glycylamino)penicillanic Acid (Ampicillin) by Manganese(III) in Pyrophosphate Medium

Subrahmanyam E., Ishwar Bhat K., Sherigara B., Satyanarayana D.

Polish Journal of Chemistry

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2000

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Vol. 74, nr 6

793-800

EN

Manganese(III) has been stabilized in weakly acidic solution by pyrophosphate and the compklex Formed was elucidated spectrophotoetrically. Stoichiometry of magnese (III) oxidation of apicillin in pyrophosphate medium was establishesd in the pH range 2.0-3.0 by iodometric and spectrophotometric methods.The oxidation follows first order kinetics in [Mn(III)]. The effect of varying [Mn(III)], [Amp], [H+], added [Mn(II)], total[P2O7 4-] and added[ClO-4],[SO4 2-] ions have been studied. The order in [Amp] is unity and increase in [H=] increases the rate. With increase in [Mn(II)] and [P2O4- 7], a retardation effect was noticed. Oxidation products have been proposed. Dependence of reaction rate on temperature has been studied and activation parameters were computed. A mechanism consistent with the observed results has been proposed.

8

Reaction of ampicillin and amoxicillin with alcohols

Frańska M.

Ars Separatoria Acta

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2012/2013

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No. 9/10

25--35

EN

HPLC/ESI-MS analyses of alcohol solutions of ampicillin and amoxicillin have shown that the two drugs react very readily with alcohols (methanol and ethanol) at room temperatures, without the presence of any catalyst. Products of methanolysis were detected even when methanol was used as a mobile phase, in HPLC/ESI-MS analysis of a water solution of ampicillin and amoxicillin. Therefore, neither methanol nor other alcohols should be used when working with these compounds.